In primary airway epithelial cells, the unjamming transition is distinct from the epithelial-to-mesenchymal transition

The epithelial-to-mesenchymal transition (EMT) and the unjamming transition (UJT) each comprises a gateway to cellular migration, plasticity and remodeling, but the extent to which these core programs are distinct, overlapping, or identical has remained undefined. Here, we triggered partial EMT (pEMT) or UJT in differentiated primary human bronchial epithelial cells. After triggering UJT, cell-cell junctions, apico-basal polarity, and barrier function remain intact, cells elongate and align into cooperative migratory packs, and mesenchymal markers of EMT remain unapparent. After triggering pEMT these and other metrics of UJT versus pEMT diverge. A computational model attributes effects of pEMT mainly to diminished junctional tension but attributes those of UJT mainly to augmented cellular propulsion. Through the actions of UJT and pEMT working independently, sequentially, or interactively, those tissues that are subject to development, injury, or disease become endowed with rich mechanisms for cellular migration, plasticity, self-repair, and regeneration.


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Policy information about availability of data All manuscripts must include a data availability statement. This statement should provide the following information, where applicable: -Accession codes, unique identifiers, or web links for publicly available datasets -A list of figures that have associated raw data -A description of any restrictions on data availability Jin-Ah Park Aug 6, 2020 Phase and fluorescent images were collected using ZEN blue (v. 2.0) software. Cell shape data was collected using SeedWater Segmenter software (v0.5.7.1). Western blot and mRNA data were collected as described in the methods section.
Custom code written in Matlab (R2019a) was used to determine dynamic and structural data from phase and fluorescent images. Cellular trajectories were generated using the optical flow (farneback) functions built into Matlab (R2019a). Images were processed in Zen blue (v. 2.0.) and ImageJ (v 1.52n). All software is available upon request.
Data that comprise the graphs within this manuscript are included in the Source Data file. Any findings or raw data of this study are available from the corresponding author upon request.

nature research | reporting summary
October 2018 Field-specific reporting Please select the one below that is the best fit for your research. If you are not sure, read the appropriate sections before making your selection. . All studies were performed with at least n=3 independent experimental replications. These final studies were carried out after extensive preliminary data was collected.
Dynamic analysis for certain data points was excluded in the case when there was extreme drift in the sample on the microscope which could not be corrected by standard drift correction methods. These were excluded before the analysis was performed, based on the observation of the extreme drift. This occurred rarely and randomly.
We use primary airway epithelial cells from non-diseased patients. We replicated our studies in cells derived from at least 3-4 donors, and the data presented in the manuscript are taken in toto from all of these experiments.
In all experiments, internal controls were present for each donor, and we present results across donors. Wells were assigned to control, compression, or TGF-beta-1 treatment groups randomly. To control for our mechanical compression condition, we exposed the control cells to a sham pressure setup. To control for our exposure to TGF-beta-1, we exposed the control cells to vehicle.
Where possible, researchers were blinded. In all cases, the same analysis process was applied to all samples, regardless of blinding. Researchers were blinded when isolating mRNA and when measuring the dynamical (speed, diffusivity, dynamic pack size) and structural (aspect ratio, orientation pack size) properties of the samples. All antibodies were used according to validation on the manufacturer's website. Western blotting: The antibody against E-cadherin was purchased from Cell Signaling Technology (CST), which validated the Ab for human Ecadherin by WB. Using this antibody at the dilution ratio recommended by CST, and at lower dilutions, we detected E-cadherin protein at the expected MW (135kDa) using human cell lysates. The antibody against N-cadherin was purchased from Cell Signaling Technology (CST), which validated the Ab for human Ncadherin by WB. Using this antibody at the dilution ratio recommended by CST (1:1000), we detected N-cadherin protein at the expected MW (140kDa) using human cell lysates. The antibody against vimentin was purchased from CST, which validated the Ab for human vimentin by WB. Using this antibody